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miR-30a靶标的生物信息学预测及其通过上调PTEN表达抑制骨肉瘤细胞增殖

Bioinformatics prediction of miR-30a targets and its inhibition of cell proliferation of osteosarcoma by up-regulating the expression of PTEN.

作者信息

Zhong Biao, Guo Shang, Zhang Wei, Zhang Chi, Wang Yukai, Zhang Changqing

机构信息

Department of Orthopedics, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, 200233, China.

出版信息

BMC Med Genomics. 2017 Nov 15;10(1):64. doi: 10.1186/s12920-017-0300-3.

Abstract

BACKGROUND

MiRNAs are frequently abnormally expressed in the progression of human osteosarcoma. Phosphatase and tensin homologue deleted on chromosome 10 (PTEN) is one of the tumor suppressors in various types of human cancer. In the present study, we detected how hsa-miR-30a-3p regulated PTEN and further tested the role of hsa-miR-30a-3p in the cell proliferation of osteosarcoma cells.

METHODS

The levels of miR-30a were determined by real time PCR. The expression of PTEN was tested by western blotting analysis. Cell distribution of PTEN was observed with confocal laser scanning microscope. Cell viability was determined by MTT assay.

RESULTS

The expression of miR-30a and PTEN was obviously decreased in MG-63, 143B and Saos-2 cells compared with primary osteoblasts. TargetScan analysis data showed miR-30a might bind with position 30-57 of 3'UTR of PTEN. Transfection with miR-30a-3p increased the level of PTEN in MG-63 cells, while transfection with miR-30a-3p inhibitor significantly decreased the expression of PTEN in osteosarcoma cells. Transfection with miR-30a-3p significantly inhibited cell proliferation of osteosarcoma cells, while miR-30a inhibitor obviously promoted cell viability of MG63 cells and Saos-2 cells. Inhibition of PTEN eliminated the proliferation inhibitory effect of miR-30a-3p.

CONCLUSION

Thus, all these findings revealed the anti-tumor effects of miR-30a in human osteosarcoma cells, which could be mediated by regulating the level of PTEN.

摘要

背景

微小RNA(miRNAs)在人类骨肉瘤进展过程中常异常表达。10号染色体缺失的磷酸酶和张力蛋白同源物(PTEN)是多种人类癌症中的肿瘤抑制因子之一。在本研究中,我们检测了hsa-miR-30a-3p如何调控PTEN,并进一步测试了hsa-miR-30a-3p在骨肉瘤细胞增殖中的作用。

方法

通过实时聚合酶链反应(PCR)测定miR-30a的水平。通过蛋白质印迹分析检测PTEN的表达。用共聚焦激光扫描显微镜观察PTEN的细胞分布。通过MTT法测定细胞活力。

结果

与原代成骨细胞相比,MG-63、143B和Saos-2细胞中miR-30a和PTEN的表达明显降低。TargetScan分析数据显示miR-30a可能与PTEN 3'非翻译区(UTR)的30-57位结合。用miR-30a-3p转染可增加MG-63细胞中PTEN的水平(表达量),而用miR-30a-3p抑制剂转染则显著降低骨肉瘤细胞中PTEN的表达。用miR-30a-3p转染可显著抑制骨肉瘤细胞的增殖,而miR-30a抑制剂则明显促进MG63细胞和Saos-2细胞的活力。抑制PTEN可消除miR-30a-3p的增殖抑制作用。

结论

因此,所有这些发现揭示了miR-30a在人类骨肉瘤细胞中的抗肿瘤作用,这可能是通过调节PTEN的水平来介导的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7ea3/5688649/72e9703e807d/12920_2017_300_Fig1_HTML.jpg

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